In vitro assay of alkaline phosphatase enzyme activity in liver and fasciola hepatica parasite

Abstract

The present work was designed to determine alkaline phosphatase (ALP) activity level in Fasciola hepatica parasite and Fasciola infected livers to evaluate the effects of liver infection on enzyme activity and to compare enzyme activity in liver and parasite. The sheep livers were collected and adult Fasciola hepatica parasites were isolated and washed with PBS buffer. Collected healthy and infected livers and parasite were homogenized and extract solutions were centrifuged and stored at -20°C. ALP enzymes activity was measured in the extract solution of samples. Proteins of the samples were measured and protein bands were detected through using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) methods respectively. Independent two samples t-test was performed to determine significant difference between different groups. The mean values of the assayed ALP specific activities of infected and healthy livers and Fasciola hepatica parasite were estimated 0.163,0.133,0.048 U/mg protein/ml correspondingly. Gel electrophoresis (SDS-PAGE) of Fasciola hepatica and liver extract solution samples displayed different protein bands, including ALP enzyme. Statistical analysis did not show significant difference between enzyme’s activity in infected and healthy livers (P>0.05). However, the liver demonstrated more than ALP activity level to parasite (P<0.05). The findings show that ALP enzymes activity in Fasciola hepatica infected livers could not be concerned as a specific pathological biomarker in fascioliasis, but meanwhile this enzyme displays interest activity in parasite.